Cigarette smoke aggravates sertoli/spermatogenic cells damage by activating the ROS/NLRP3 signaling pathway
10.19405/j.cnki.issn1000-1492.2023.09.020
- Author:
Shuomin Wang
1
;
Yifan Hu
1
;
Bingxian Zhou
1
;
Shiheng Fan
1
;
Qisheng Wu
1
;
Yayan Fan
1
;
Chunyun Lu
1
;
Dee Yu
1
;
Jing Zhang
1
Author Information
1. International School of Public Health and One Health , Hainan Medical University, Haikou 571199
- Publication Type:Journal Article
- Keywords:
cigarette smoke;
NLRP3;
ROS;
pyroptosis;
sertoli/spermatogenic cells
- From:
Acta Universitatis Medicinalis Anhui
2023;58(9):1557-1564
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To investigate the mechanism of sertoli/spermatogenic injury induced by cigarette smoke extract(CSE) based on ROS/NLRP3 signaling pathway.
Methods :The sertoli/spermatogenic of rat were cultured in vitro , and they were divided into control group , cigarette Smoke extract (CSE) group and CSE + NLRP3 inflammosome inhibitor MCC950 group. Cell viability was detected by CCK⁃8 assay; Lactate dehydrogenase ( LDH) was used to detect cell membrane damage ; reactive oxygen species ( ROS) production was used to detect oxidative stress level ; Hoechst/PI fluorescent staining was used to detect cell pyroptosis. Real⁃time quantitative PCR ( RT⁃qPCR) was used to determine the mRNA level of NLRP3 , ASC , Caspase⁃1 , GSDMD , IL⁃1β and IL⁃18. Western blot was used to determine the protein level of NLRP3 , ASC , Caspase⁃1 , GSDMD , IL⁃1β and IL⁃18.
Results:CCK⁃8 assay showed that cell viability decreased in CSE group (P < 0. 001) , LDH leakage rate (P < 0. 001) and ROS (P < 0. 001) production increased. Compared to the CSE group , cell viability increased (P < 0. 001) , LDH leakage rate (P < 0. 01) and ROS (P < 0. 001) production decreased in CSE + MCC950 group. The number of PI staining cells in CSE group was more than that in control group and CSE + MCC950 group , but staining cells in CSE + MCC950 group was less than that in CSE group. RT⁃qPCR and Western blot results showed that the expression levels of NLRP3 , ASC , Caspase⁃1 , GSDMD , IL⁃1β and IL⁃18 increased in CSE group but decreased in CSE+ MCC950 group.
Conclusion :The oxidative stress and inflammation of sertoli/spermatogenic induced by ciga⁃ rette smoke may be related to the activation of ROS/NLRP3 signaling pathway.
- Full text:2024081216365614166烟草烟雾通过激活ROS_N...toli-生精细胞损伤研究_王硕敏.pdf
